We continue to develop our existing biological atomic force (Bio-AFM) platforms, Quartz Crystal Microbalance-Dissipation (QCM-D), super-resolution optical microscopy, and related measurement systems, and to apply these biophysical technologies to a number of biomedical and biological investigations in collaboration with NIH intramural and extramural researchers. Major collaborations with notable results include: (A) We have investigated the macromolecular structure and nanomechanical properties of many malaria vaccine candidates via Bio-AFM and related bioanalysis in collaboration with Dr. David Narum (NIAID, NIH), Dr. Patrick Duffy (NIAID, NIH) and other co-investigators at the Laboratory of Malaria Immunology and Vaccinology, NIAID, NIH. These protein antigens for malaria vaccine are being produced via recombinant-protein biotechnology, purified, and characterized in a manner suitable for human trials and scale-up productions. We have focused on using AFM and QCM-D to understand the structural properties of the developing vaccine products, including Qbeta virus-like particles as developing malaria vaccine carriers for enhanced immunological response, and are preparing manuscripts for publications. (B) We have continued our Bio-AFM studies of clathrin and clathrin coated vesicles (CCVs) in collaboration with Dr. Ralph Nossal (NICHD, NIH), Prof. Eileen Lafer (Univ. Texas Health Sciences Center, San Antonio), and others. Clathrin triskelia form the outer clathrin lattice cages of the CCVs during subcellular trafficking via interactions with adaptor proteins, membrane lipids, and other cofactors. The intricacies of the clathrin-mediated dynamic macromolecular constructs have inspired numerous structural and functional studies, including several publications from this collaboration. We have used systematic QCM-D measurements to enhance the accuracy of the nanomechnical characterization for clathrin, CCVs, and other assembled structures. A new manuscript is nearing submission. (C) We have advanced a number of new intramural collaborations involving Bio-AFM and related nanotechnologies. Two new collaborations have yielded recent publications: (1) with Dr. Ashwin Bhirde (LOMIN, NIBIB), Dr. Xiaoyuan Chen (LOMIN, NIBIB) and other co-investigators on carbon nanotubes and live cells, and (2) with Ms. Katelyn Nagy (CBL, MDP, NCI), Dr. Joel Schneider (CBL, MDP, NCI) and coworkers on a new class of protein biopolymer hydrogels. More results from such new efforts, as well as ongoing older collaborations, are forthcoming.